Antioxidant micronutrients in electronic cigarettes

ABSTRACT

An antioxidant formulation for use with electronic cigarettes is provided and the formulation comprises: Vitamin A, Trans-Resveratrol, polyphenol, and tertiary butyl hydroquinone

RELATED APPLICATION

This is a continuation-in-part of U.S. application Ser. No. 14/756,311entitled Antioxidant Micronutrients Formulation in ElectronicCigarettes.

BACKGROUND OF THE INVENTION Field of the Invention

This invention presents novel concepts relative to the use of uniqueantioxidant strategies to reduce the potential adverse effects ofelectronic cigarette (E-cig) use. The strategies include decreasingoxidative damage and inflammation, major factors in E-cig relatedadverse health effects, by employing agents that 1) directly scavengeexcess free radicals by elevating dietary and endogenous antioxidantlevels, and 2) indirectly reduce oxidative damage and inflammation byactivating factors that increase antioxidant enzyme levels. The strategyalso employs a novel mixture of agents that are incorporated directlyinto the E-cig liquid as well as an orally-consumed unique combinationof antioxidant micronutrients.

SUMMARY OF THE INVENTION

In an embodiment, the present invention provides for an antioxidantformulation for supporting health in human beings comprising: Vitamin A,Vitamin E, Trans-Resveratrol, polyphenol, and tertiary butylhydroquinone (TBHQ). In another embodiment, the present inventionrelates to an antioxidant formulation is to be used in a liquid form forelectronic cigarettes. In yet another embodiment, the present inventionprovides for an antioxidant formulation is composed of lipid solublecomponents. In a further embodiment, the present invention relates to anantioxidant formulation is to be inhaled by human beings. In stillanother embodiment, the present invention provides for an antioxidantformulation wherein additional ingredients are chosen from a groupcomprising: Vitamin A, Vitamin E, Vitamin C, Calcium, Niacin, Vitamin D,Vitamin B, Folic Acid, Biotin, Pantothenic Acid, Co-enzyme Q10, N-acetylcysteine, and Alpha Lipoic Acid. In still yet another embodiment, thepresent invention provides for an antioxidant formulation is to besupplemented with a secondary antioxidant formulation. In a futherembodiment, the present invention provides for a secondary antioxidantformulation comprises Vitamin A, Vitamin E, Natural mixed carotenoids,Vitamin C, Vitamin D, B-vitamins, Selenium, Co-enzyme Q10, Alpha-lipoicacid, N acetyl cysteine, L-carnitine, Omega-3-fatty acids, Resveratrol,Green tea polyphenol, Curcumin, Genistein, and Allicin. In anotherembodiment, the present invention provides for a secondary antioxidantformulation is to be taken orally. In still a further embodiment, thepresent invention relates to secondary antioxidant formulation is to betaken at least once a day. In another embodiment, the present inventionprovides for secondary antioxidant formulation of is to be taken twice aday.

In a further embodiment, an e-cigarette cartridge containing anatomizable liquid composition comprising Vitamin A, trans-resveratrol, agreen tea polyphenol, and tert-butylhydroquinone, wherein the atomizableliquid composition is nicotine free.

In yet another embodiment, wherein the Vitamin A is retinyl palmitate.

In still another embodiment, an e-cigarette cartridge wherein the amountof retinyl palmitate in the atomizable liquid composition is from 20 IUto 3,000 IU.

In yet a further embodiment, an e-cigarette cartridge wherein the amountof trans-resveratrol in the atomizable liquid composition is from 1 mgto 100 mg.

In a further embodiment, an e-cigarette cartridge wherein the amount ofgreen tea polyphenol in the atomizable liquid composition is from 1 mgto 100 mg.

In still a further embodiment, an e-cigarette cartridge wherein theamount of tert-butylhydroquinone in the atomizable liquid composition isfrom 0.5 mg to 20 mg.

In yet a further embodiment, an e-cigarette cartridge containing anatomizable liquid composition comprising: retinyl palmitate;trans-resveratrol; a green tea polyphenol; and tert-butylhydroquinone;and wherein the atomizable liquid composition is nicotine free.

In another embodiment, an e-cigarette cartridge of claim 33, wherein theamount of retinyl palmitate in the atomizable liquid composition is from20 IU to 3,000 IU.

In yet another embodiment, an e-cigarette cartridge wherein the amountof trans-resveratrol in the atomizable liquid composition is from 1 mgto 100 mg.

In still another embodiment, an e-cigarette cartridge wherein the amountof green tea polyphenol in the atomizable liquid composition is from 1mg to 100 mg.

In another embodiment, an e-cigarette cartridge wherein the amount oftert-butylhydroquinone in the atomizable liquid composition is from 0.5mg to 20 mg.

In a further embodiment, an e-cigarette cartridge containing anatomizable liquid composition comprising: retinyl palmitate, wherein theamount of retinyl palmitate in the atomizable liquid composition is from20 IU to 3,000 IU; trans-resveratrol, wherein the amount oftrans-resveratrol in the atomizable liquid composition is from 1 mg to100 mg; a green tea polyphenol; and tert-butylhydroquinone, wherein theamount of tert-butylhydroquinone in the atomizable liquid composition isfrom 0.5 mg to 20 mg; and wherein the atomizable liquid composition isnicotine free.

In still a further embodiment, the e-cigarette cartridge of claim 40,wherein the amount of green tea polyphenol in the atomizable liquidcomposition is from 1 mg to 100 mg.

DETAILED DESCRIPTION OF THE INVENTION

As required, detailed embodiments of the present invention are disclosedherein; however, it is to be understood that the disclosed embodimentsare merely exemplary of the invention that may be embodied in variousforms. The figures are not necessarily to scale, some features may beexaggerated to show details of particular components. Therefore,specific structural and functional details disclosed herein are not tobe interpreted as limiting, but merely as a basis for the claims and asa representative basis for teaching one skilled in the art to variouslyemploy the present invention.

The specific example below will enable the invention to be betterunderstood. However, they are given merely by way of guidance and do notimply any limitation.

Because tobacco cigarette smoking caused adverse health effects andcessation was difficult due to nicotine addiction, efforts were made todevelop a non-tobacco cigarette. In 1963, Dr. Herbert A. Gilbertpatented a smokeless non-tobacco cigarette which drew moistened airthrough a heating element to generate vapor resembling tobacco cigarettesmoke. In 2003, Hon Lik, a Chinese pharmacist, patented anElectronic-cigarette (E-cig) which was essentially a rechargeablelithium battery powered device that produced an aerosol vapor resemblingtobacco cigarette smoke. The vapor is formed by heating a solution ofpropylene glycol and glycerin with or without nicotine at hightemperatures (40-65° C.). Whether E-cigs have less long-term healthconsequences than tobacco cigarettes or aid in smoking cessation isunknown. Limited studies have been performed on the effects of vapor onhuman health but a comparative analysis of E-cig vapor and tobaccocigarette smoke showed that some common toxic chemical levels are lowerin E-cigs. Some additional toxic chemicals that are found in the vaporof E-cigs that are not present in tobacco cigarette smoke, and viceversa.

It is proposed that increased oxidative stress and chronic inflammationproduced by the constituents of the E-cig vapor are primary biochemicaldefects that may contribute to adverse health effects. Therefore, thisinvention describes the scientific rationale and evidence to suggestthat addition of multiple micronutrients into the E-cig liquid mayprotect against injurious effects of toxic chemicals in the vapor. Inaddition, an oral supplement of micronutrients may provide overallprotection against oxidative damage and chronic inflammation produced bythe oxidizing agents, carcinogens and ultrathin particles of the heavymetals in the vapor.

E-cig Components:

An E-cig consists of a stainless steel shell, a rechargeable lithium ionbattery, a small chamber containing miniaturized electronics and a tinyatomizer which vaporizes the E-cig liquid that may contain propyleneglycol, glycerin, with or without varying amounts of nicotine andflavoring which include tobacco, menthol, mint, chocolate, coffee,apple, cherry, and caramel. The E-cig liquid fills the cartridge. Thebattery is connected to the vaporizer chamber which contains electroniccontrol chips and an atomizer. The atomizer consists of a small heatingelement which vaporizes the E-cig liquid, as well as wicking materialsthat draw in the liquid. The atomizer and cartridge are also combined ina single unit called a cartomizer. Many E-cigs have a light-emittingdiode on the end which lights up resembling actual flame when the usersinhale. The user attaches a cartridge containing the E-cig liquid to thevaporizing chamber. The tip of the cartridge serves as the mouth pieceand the user inhales the way they would inhale a tobacco cigarette. Thisinhalation process activates the atomizer to heat the E-cig liquidcontaining nicotine in the cartridge in order to produce vaporresembling tobacco cigarette smoke during exhalation. The nicotine vaporenters the lung and produces a nicotine “high”.

Chemical Constituents of E-cig Vapor:

The levels of chemical constituents such as nicotine, toxic organiccompounds and heavy metals in the E-cig vapor are highly variable (1-4).Although E-cigs deliver nicotine at a level of about 20% of thatdelivered by tobacco cigarettes, saliva levels of nicotine are similarin both sets of users (5, 6). The levels of toxic and carcinogeniccompounds in E-cig vapor were 1-2 orders of magnitude lower than in thetobacco cigarette smoke, but higher than in nicotine inhalers (7). TheE-cig vapor also contains propylene oxide (a product of heated propyleneglycol), a potential carcinogen, acrolein (a product of heated glycerin)which can cause irritation in the upper respiratory tract and, in someproducts, detectable levels of tobacco-specific nitrosamines (8-10).Although the vapor of E-cigs may contain formaldehyde, acetaldehyde,isoprene, acetic acid,2-butanodione, acetone, propanol, propylene glycoland diacetin, the levels were lower than in tobacco cigarette smoke(11). A comparative analysis of potentially toxic and carcinogeniccompounds, such as carbonyls, volatile organic compounds, nitrosaminesand heavy metals in E-cig vapor showed levels that were many times lowerthan those in tobacco cigarette smoke (7).

Various E-cig components contain metals, such as copper wire coated withsilver, tin, copper, aluminum, nickel, and silicate. E-cig vapor maycontain particles of tin, silver, iron, copper, aluminum and silicate,and nanoparticles of tin, chromium and nickel (3). The concentrations ofthese metals in the E-cig vapor were often higher than those found intobacco cigarette smoke. These particles can induce inflammation intissues especially in the lung. Nanoparticles from the vapor canaccumulate in various tissues including the lung, liver, kidney, heartand brain (12-15).

Effects on Human Health:

The users of E-cigs develop nicotine addiction similar to that found insmokers of tobacco cigarettes. E-cig vapor with ultrathin particles oftin was very toxic to human pulmonary fibroblasts (3). Ultrathinparticles of metals produced cytotoxic effects in vitro and in vivo (16,17). Cobalt, chromium and chromium oxide nanoparticles can be toxic tomammalian cells in culture (18, 19), whereas nanoparticles of nickelhydroxide can induce oxidative stress and inflammation in the lung andheart of mice (20). Other adverse health effects of E-cigs includethroat and mouth irritation, cough, nausea, and vomiting (4). In a mousemodel, long term exposure to E-cig vapor increased infiltration ofinflammatory cells including eosinophils into the airways, stimulatedthe production of cytokines interleukin-4 (IL-4), IL-5 and IL-13, andaggravated the asthmatic airway inflammation and airwayhyper-responsiveness (21).

In clinical experience, a 20 year-old user of E-cigarettes developedacute eosinophilic pneumonia (22). In nine volunteer E-cig users, thelevels of functional exhaled nitric oxide, a quantitative measure ofairway inflammation, increased in the vapor. In addition, the levels of1, 2-propannediol, glycerin, nicotine, polycyclic aromatic hydrocarbons,carbonyls and ultrathin particulate matters of certain metals increasedin the indoor air possibly inducing a “second-hand smoke” phenomenon(23). Because of the presence of high levels of nicotine, ultrathinparticles of metals, oxidizing agents and volatile organic compounds inthe E-cig vapor, the risk of cardiac arrhythmias and hypertension mayincrease and enhance the possibility of developing cardiovascular events(24). In addition, an increased risk of lung disease also exists inE-cig users. It has been suggested that E-cig use may constrict airwaysand aggravate the symptoms of asthma, emphysema or chronic bronchitis(25). Also, the levels of exhaled nitric oxide increased in users ofE-cigarettes containing nicotine, but not in users of E-cigaretteswithout nicotine (23). Increased production of nitric oxide can lead toenhanced levels of peroxynitrite, a deadly form of free radical.

Reducing Health Effects of E-cigs by Decreasing Oxidative Stress andInflammation:

Since E-cig vapor contains oxidizing agents such as propylene oxide,carcinogens such as nitrosamines, inflammatory agents such as acrolein,and ultrathin particles of heavy metals, it is likely that long-termE-cig may induce increased oxidative stress and chronic inflammation invarious tissues, especially in the lung. Chronic inflammation releasespro-inflammatory cytokines, prostaglandins, adhesion molecules,complement proteins and free radicals all of which are toxic to cells.Ultraparticles of heavy metals would primarily accumulate in lung tissueand be a constant source of inflammation. Since these particles cannotbe readily removed, it would be desirable to block their effects and/orprotect tissues from the action of toxic chemicals released frominflammatory cells. Thus, reducing oxidative stress and chronicinflammation is one of the rational choices for reducing the potentialhealth risks E-cigs in humans.

Optimally Reducing Oxidative Stress and Inflammation:

Elevation of the levels of antioxidant enzymes as well as dietary andendogenous antioxidant chemicals is essential for optimally reducingoxidative stress and chronic inflammation. Supplementation withantioxidant micronutrients can elevate their levels in the body.However, elevation of the levels of antioxidant enzymes is a complexprocess. The levels of antioxidant enzymes are elevated by activation ofa transcriptional factor nuclear factor-erythroid 2-related factor -2(Nrf2) which translocates itself from the cytoplasm to the nucleus whereit binds with the antioxidant response element (ARE) to increase thetranscription of genes coding for antioxidant enzymes. Activation ofNrf2 occurs by reactive oxygen species (ROS)-dependent andROS-independent mechanisms. In addition, elevated levels of antioxidantenzymes are also dependent upon the binding ability of Nrf2 with ARE inthe nucleus. The age-related decline in antioxidant enzymes in the liverof old rats compared to that in young rats was due to this reduction inthe binding ability of Nrf2 with ARE. Treatment with alpha-lipoic acidrestored this defect, increased the levels of antioxidant enzymes andrestored the loss of glutathione in the liver of old rats (26).

ROS-dependent mechanism of Nrf2 activation: Normally, Nrf2 is associateda protein which acts as an inhibitor of Nrf2 (INrf2) in the cytoplasm(27). INrf2 acts as a sensor for ROS/electrophilic stress. In responseto increased ROS, Nrf2 dissociates itself from INrf2 and translocatesinto the nucleus where it binds with ARE and up-regulates antioxidantgenes.

ROS-independent mechanism of Nrf2 activation: Antioxidants such asvitamin E, the flavonoid, genistein (28), allicin, an organosulfurcompound in garlic (29), sulforaphane, a organosulfur compound incruciferous vegetables (30), kavalactones such as methysticin, kavainand yangonin (31) and dietary restriction (32) activate Nrf2 byROS-independent mechanisms.

Nrf2 Response to ROS during Acute and Chronic Oxidative Stress:

Acute oxidative stress during strenuous exercise causes translocation ofNrf2 from the cytoplasm to the nucleus where it binds with ARE toup-regulate antioxidant genes. However, during chronic oxidative stressNrf2 becomes unresponsive to ROS. Therefore, supplementation withselected non-toxic agents in combination may be useful in optimallyreducing chronic oxidative stress because they can activate Nrf2 byROS-independent mechanisms as well as elevate the levels of dietary andendogenous antioxidant micronutrients.

1. Agents that reduce oxidative stress by directly scavenging freeradicals: These include dietary antioxidants, such as vitamin A,beta-carotene, vitamin C, and vitamin E, and endogenous antioxidants,such as glutathione, alpha-lipoic acid, and coenzyme Q10.2. Agents that reduce oxidative stress by activating Nrf2 via anROS-independent mechanism: These include vitamin E (28) and coenzyme Q10(33).3. Agents that reduce oxidative stress directly by scavenging freeradicals as well as indirectly by activating Nrf2 via an ROS-independentmechanism: These include vitamin E (28), alpha-lipoic acid,(26),curcumin (34), resveratrol (35, 36), omega-3-fatty acids (37, 38) andN-acetyl cysteine (NAC) (39).

Reducing Chronic Inflammation:

Activation of Nrf2 suppresses inflammation. Antioxidants and otheragents, individually and in combination, from the above groups have beenshown to reduce chronic inflammation (40-47).

Example Antioxidant Formulation for E-cig Liquid: Primary Lipid SolubleComponents:

Per ml of liquid (approximately equivalent to smoking two packs oftobacco cigarettes):

Target Dose: Dosage Range: Vitamin A (retinyl palmitate) 2000 IU 20-3000IUVitamin E (d-alpha-tocopheryl acetate) 25 IU 0.5-50 IU Vitamin E(d-alpha-tocopheryl succinate) 25 IU 0.5-50 IU Trans-Resveratrol 20 mg1-100 mg Green tea polyphenol 20 mg 1-100 mg Tertiary butyl hydroquinone(TBHQ) 2 mg 0.5-20 mg

Doses in the above example are recommended but can be flexible. Thesynthetic antioxidant TBHQ is included in the E-cig liquid to preventformation of oxidation products of propylene glycol and otherconstituents and, thereby, reduce the damaging effects of oxidizedproducts on the lung. In addition, a synthetic antioxidant is necessaryto prevent the degradation of vitamins A and E in order to increase thelevels of antioxidants in the lung. The primary lipid solublecombination can optimally reduce oxidative stress and chronicinflammation in the lung indirectly by activating the Nrf2/ARE pathwayby an ROS-independent mechanism, and directly by scavenging freeradicals.

Example Formulation of Secondary Components:

-   Vitamin A 29.41 IU-   Vitamin C 5.88 mg-   Vitamin E (d-alpha tocopheryl acetate) 1.18 IU-   Vitamin E (d-alpha tocopheryl succinate) 1.18 IU-   Calcium 0.62 mg-   Niacin 0.18 mg-   Vitamin D3 2.35 IU-   Vitamin B1 0.024 mg-   Vitamin B2 0.03 mg-   Vitamin B6 0.03 mg-   Folic Acid 4.7 mcg-   Vitamin B12 0.06 mcg-   Biotin 1.18 mcg-   Pantothenic Acid 0.06 mg-   Co-enzyme Q10 0.22 mg-   N-acetyl cysteine 1.47 mg-   Alpha Lipoic Acid 0.22 mg    Doses in the above example are recommended but can be flexible.

Temperature Stability:

A temperature range of 45-60° C. is needed to convert E-cig liquid intovapor. All polyphenols including resveratrol and green tea polyphenolare stable at a temperature 100° C. At 125° C., significant degradationof these polyphenols occurs (48). Degradation of vitamin E occurs at atemperature range of 180-260° C. for 20-80 minutes. This can beprevented by TBHQ (49). Vitamin A levels during sterilization ofpre-pasteurized milk at 85° C. were maintained above 95% (50).

Supplemental Oral Micronutrients:

Oxidizing agents, carcinogens and ultraparticles of metals are presentin E-cig vapor and not only accumulate in the lung, but also in theblood stream and are deposited in other organs including the brain. Inorder to protect the organ systems of the entire body, it is essentialto supplement the antioxidants in the E-cig liquid with orally-consumedmicronutrients.

Example Daily Formulation for E-cig Users

-   Vitamin A (retinyl palmitate)-   Vitamin E (both d- alpha-tocopherol and d-alpha-tocopheryl    succinate)-   Natural mixed carotenoids,-   Vitamin C (calcium ascorbate)-   Vitamin D-   B-vitamins,-   Selenium-   Co-enzyme Q10-   Alpha-lipoic acid-   N acetyl cysteine-   L-carnitine-   Omega-3-fatty acids-   Resveratrol-   Green tea polyphenol-   Curcumin-   Genistein-   Allicin

Doses in the above example are recommended but can be flexible. Theseagents can optimally reduce oxidative stress and chronic inflammationindirectly by activating the Nrf2/ARE pathway by an ROS-independentmechanism, and by directly scavenging free radicals. No iron, copper ormanganese is included because these trace minerals are known to interactwith vitamin C to produce free radicals. These trace minerals areabsorbed from the intestinal tract more readily in the presence ofantioxidants than in their absence and that could result in excess bodystores of these minerals. Increased iron stores have been linked toincreased risk of several chronic diseases (51).

The supplement combination should be taken orally and divided into twodoses, half in the morning and the other half in the evening with ameal. This is because the biological half-lives of micronutrients arehighly variable which can create excessive fluctuations in the tissuelevels of these micronutrients. Even only a two-fold difference in thelevels of certain micronutrients such as alpha-tocopheryl succinate cancause a marked difference in the expression of gene profiles (Prasad,KN, unpublished data). The twice-daily dosing will help maintainrelatively consistent levels of micronutrients in the body.

Numerous modifications and variations of the present invention arepossible in light of the above teachings. It is therefore to beunderstood that within the scope of the attendant claims attachedhereto, this invention may be practiced otherwise than as specificallydisclosed herein.

What is claimed is:
 1. An e-cigarette cartridge containing an atomizableliquid composition comprising Vitamin A, trans-resveratrol, a green teapolyphenol, and tert-butylhydroquinone, wherein the atomizable liquidcomposition is nicotine free.
 2. The e-cigarette cartridge of claim 1,wherein the Vitamin A is retinyl palmitate.
 3. The e-cigarette cartridgeof claim 2, wherein the amount of retinyl palmitate in the atomizableliquid composition is from 20 IU to 3,000 IU.
 4. The e-cigarettecartridge of claim 1, wherein the amount of trans-resveratrol in theatomizable liquid composition is from 1 mg to 100 mg.
 5. The e-cigarettecartridge of claim 1, wherein the amount of green tea polyphenol in theatomizable liquid composition is from 1 mg to 100 mg.
 6. The e-cigarettecartridge of claim 1, wherein the amount of tert-butylhydroquinone inthe atomizable liquid composition is from 0.5 mg to 20 mg.
 7. Ane-cigarette cartridge containing an atomizable liquid compositioncomprising: retinyl palmitate; trans-resveratrol; a green teapolyphenol; and tert-butylhydroquinone; and wherein the atomizableliquid composition is nicotine free.
 8. The e-cigarette cartridge ofclaim 7, wherein the amount of retinyl palmitate in the atomizableliquid composition is from 20 IU to 3,000 IU.
 9. The e-cigarettecartridge of claim 7, wherein the amount of trans-resveratrol in theatomizable liquid composition is from 1 mg to 100 mg.
 10. Thee-cigarette cartridge of claim 7, wherein the amount of green teapolyphenol in the atomizable liquid composition is from 1 mg to 100 mg.11. The e-cigarette cartridge of claim 7, wherein the amount oftert-butylhydroquinone in the atomizable liquid composition is from 0.5mg to 20 mg.
 12. An e-cigarette cartridge containing an atomizableliquid composition comprising: retinyl palmitate, wherein the amount ofretinyl palmitate in the atomizable liquid composition is from 20 IU to3,000 IU; trans-resveratrol, wherein the amount of trans-resveratrol inthe atomizable liquid composition is from 1 mg to 100 mg; a green teapolyphenol; and tert-butylhydroquinone, wherein the amount oftert-butylhydroquinone in the atomizable liquid composition is from 0.5mg to 20 mg; and wherein the atomizable liquid composition is nicotinefree.
 13. The e-cigarette cartridge of claim 12, wherein the amount ofgreen tea polyphenol in the atomizable liquid composition is from 1 mgto 100 mg.